Intracellular ROS were detected using the 2,7-dichlorodihydrofluoresceindiacetate (DCFH-DA) ROS probe (Beyotime Institute of Biotechnology, China). Briefly, H9c2 cells were incubated with the probe (10 μM) for 30 min at 37°C in the dark. Measurements of the fluorescence were performed by FACS Calibur Flow Cytometer (BD Biosciences, San Jose, CA).
Dcfh da ros probe
Manufactured by Beyotime
Sourced in China
DCFH-DA (2',7'-Dichlorodihydrofluorescein diacetate) is a fluorogenic probe commonly used for the detection of intracellular reactive oxygen species (ROS). It is a non-fluorescent, cell-permeable compound that, upon oxidation by ROS, is converted to the highly fluorescent 2',7'-dichlorofluorescein (DCF). The intensity of the fluorescent signal is proportional to the level of ROS present in the sample.
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2 protocols using dcfh da ros probe
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Intracellular ROS Detection by DCFH-DA
2
Intracellular ROS Measurement in HT22 Cells
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The HT22 cells were seeded in 96-well plates at a density of 4 × 103 cells per well. The 2,7-dichlorodihydrofluorescence indiacetate (DCFH-DA) ROS probe (Beyotime Biotechnology, Shanghai, China) was used to detect intracellular ROS. The cells were incubated with 20 μM DCFH-DA probe at 37 °C for 30 min in the dark and then washed with PBS three times. The stained cells were reviewed under a microscope, and images were captured via the use of a digital scanning microscopy imaging system (PreciPoint, Freising, Germany).
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